FIG 7.

HCV NS3 attenuates the inhibitory effect of TRIB3 on Akt phosphorylation and prevents ER stress-mediated apoptosis. (A) Huh7 cells were transfected with either vector- or Myc-tagged NS3. Twenty-four hours after transfection, cells were treated with either dimethyl sulfoxide (DMSO) (−) or 2 μg/ml of tunicamycin. Eighteen hours after treatment, total cell lysates were immunoblotted with the indicated antibodies. Protein band intensities of p-Akt/Akt and TRIB3/GAPDH were analyzed by using ImageJ. (B) Huh7 cells were transfected with either the vector or Flag-tagged TRIB3 in the absence or presence of Myc-tagged NS3. Twenty-four hours after transfection, cells were treated with either dimethyl sulfoxide (−) or tunicamycin, as indicated. Eighteen hours after treatment, total cell lysates were immunoblotted with the indicated antibodies. Protein band intensities of p-Akt/Akt were determined by using ImageJ. (C) Huh7 cells were transfected with Flag-tagged TRIB3 in the absence or presence of Myc-tagged NS3. Twenty-four hours after transfection, cells were either left untreated or treated with tunicamycin. Forty-eight hours after treatment, the percentage of adherent cells was measured by staining with crystal violet, and the percentage of dead cells was then determined.